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Pressurized liquid extraction prior to liquid chromatography with electrochemical detection for the analysis of vitamin E isomers in seeds and nuts.

Pressurized liquid extraction (PLE) was used to isolate tocopherols from seeds and nuts. Very clean extracts were obtained, which were injected directly into the chromatographic system. This enables rapid and simple control in food analysis. The PLE extraction temperature was set at 50 degrees C, with two cycles of extraction, a static time of 5 min, and acetonitrile as the extraction solvent. LC separation was accomplished on a Synergi Hydro-RP column with methanol-water (99.9:0.1, v/v) containing 2.5 mM acetic acid/sodium acetate buffer, as eluent. Coulometric detection, with a porous graphite electrode at +700 mV, was used. The method was successfully applied to the determination of alpha-, (beta + gamma)- and delta-tocopherols in almonds, sunflower seeds, hazelnuts and walnuts. The recoveries were in the 82-110% range. The results were validated with those obtained using sample treatment including alkaline hydrolysis.

Chromatography, Liquid↗

Food consumption patterns in an affluent society and barriers to overcoming dietary change.

In affluent nations, there has been an increasing awareness of the links between diet and the incidence of chronic diseases such as heart disease, hypertension and cancer. In some of these countries, this has led to improvements in the dietary profiles of the community and its health-related status. Studies of random samples of the Australian population carried out in the last decade or so have shown a downward trend in the consumption of total and saturated fats and salt and an increase in polyunsaturated fat and fibre. However, detailed assessment of food consumption patterns reveal that the changes in dietary patterns relate predominantly to improvement in choice within food categories rather than a fundamental change in the relative amounts of foods consumed across categories such as meats, dairy foods, cereal grains and fruits and vegetables. The surveys show that knowledge about the links between diet and disease is encouragingly high in most sectors of the Australian community as is concern about the healthiness of the food supply but time constraints, the perception that "healthy" foods are boring foods, lack of information on packaging and family pressures were seen as major barriers to dietary change. A further barrier is lack of knowledge about the balance of foods required and in particular, the value of cereal foods. Analysis of the dietary data show that further significant improvements in the dietary profile are only likely to accrue from a fundamental change in the relative use of the major food categories.

Attitude to Health↗

Automated food microbiology: potential for the hydrophobic grid-membrane filter.

Bacterial counts obtained on hydrophobic grid-membrane filters were comparable to conventional plate counts for Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus in homogenates from a range of foods. The wide numerical operating range of the hydrophobic grid-membrane filters allowed sequential diluting to be reduced or even eliminated, making them attractive as components in automated systems of analysis. Food debris could be rinsed completely from the unincubated hydrophobic grid-membrane filter surface without affecting the subsequent count, thus eliminating the possibility of counting food particles, a common source of error in electronic counting systems.

Bacteria↗

Spectral surface plasmon resonance biosensor for detection of staphylococcal enterotoxin B in milk.

This work evaluates a newly developed wavelength modulation-based SPR biosensor for the detection of staphylococcal enterotoxin B (SEB) in milk. Two modes of operation of the SPR biosensor are described: direct detection of SEB and sandwich assay. In the sandwich assay detection mode, secondary antibodies are bound to the already captured toxin to amplify sensor response. Samples including SEB in buffer and SEB in milk were analyzed in this work. The SPR biosensor has been shown to be capable of directly detecting concentrations of SEB in buffer as low as 5 ng/ml. In sandwich detection mode, the lowest detection limit was determined to be 0.5 ng/ml for both buffer and milk samples. The reported wavelength modulation-based SPR sensor provides a generic platform which can be tailored for detection of various foodborne pathogens and agents for food analysis and testing.

Animals↗

Immunosensors.

Immunosensors are important analytical tools for monitoring antibody-antigen reactions in real time. Recent developments in immunosensors have produced systems that allow rapid and continuous analysis of the binding event without the requirement for added reagents or separation/washing steps. As a result, great interest has focused on commercializing immunosensors for applications in areas such as clinical, environmental and food analysis.

Animals↗

Optimization of carbohydrate silylation for gas chromatography.

We developed and optimized a new carbohydrate mono- and disaccharides silylation reaction, replacing pyridine and requiring lower reaction temperature and less time. Our method consists of three basic steps. The first one is oxime formation, the second one silylate derivative and the last one gas chromatography separation and quantification with an internal standard. We evaluated several solvents, including acetonitrile, hydroxylamine and aniline. We found aniline to be the best reaction media for oxime formation with hydroxylamine hydrochloride. Among silylation agents we found N,O-bis(trimethyl)trifluoroacetamide (BSTFA) was the most efficient. Together these reagents favored both a short analysis time and fewer by-products. We evaluated the method with model solutions containing: arabinose and co-eluting xylose, fructose, glucose, sucrose and salicin (internal standard) and found it suitable for processed food analysis.

Carbohydrates↗

Method development and analysis of retail foods and beverages for carotenoid food colouring materials E160a(ii) and E160e.

An analytical method using high-performance liquid chromatography with photodiode array detection was developed and applied to the determination of the permitted food colour additives beta-carotene(E160a(ii)) and beta-apo-8'-carotenal (E160e) in foods and beverages. The scope of previously reported methods has been broadened to cover a wide range of retail foods and enzymatic hydrolysis has been used in place of saponification for high-fat samples. Quantitative results (greater then 0.1 mg kg(-1)) are given for the major colour principals trans-beta-apo-8'-carotenal and trans-beta-carotene. Semiquantitative results are given for the various cis-isomers of each colorant for which authentic reference standards were not available. The method has been used successfully for the analysis of a wide range of foodstuffs with differing fat content without the need for saponification, except for moderate- to high-fat foodstuffs containing significant levels of emulsifiers, for which it was limited. The results suggest that beta-apo-8'-carotenal (E160e) does not have widespread use in the UK. None of the samples exhibited a total beta-carotene content greater than 20 mg kg(-1) and none of the high-fat samples and only one of the 17 low-fat/beverage samples contained total beta-carotene at levels less than 0.1 mg kg(-1). The total beta-carotene contents of the low-fat/beverage samples ranged from 0.4 +/- 0.03 to 8.4 i 0.71 mg kg(-1), and the total beta-carotene contents of the high-fat samples ranged from 0.1 +/- 0.01 (jelly confectionery) to 18.5 +/- 0.98 mg kg(-1) (processed cheese).

Antioxidants↗

Assessing food system attitudes among youth: development and evaluation of attitude measures.

OBJECTIVE: To design and pilot a questionnaire for assessing youth attitudes about the food system. DESIGN: Nutrition educators provided important topics for food system education through a survey using the Delphi method. The resulting questionnaire of attitude scales was piloted with high school students. SETTING: The Delphi survey of nutrition educators was conducted through the mail and electronically. A paper and pencil questionnaire was administered to high school students in Family and Consumer Science classes. PARTICIPANTS: Delphi participants were members of the Society for Nutrition Education's Division of Sustainable Food Systems. A convenience sample of high school students completed the pilot questionnaire. VARIABLES MEASURED: Scales measured attitudes about farmland protection, participation in the local food system, hunger in the United States, organic agriculture, and environmental issues related to food. ANALYSIS: Factor analysis was used to identify constructs. The internal consistency of each resulting scale was assessed using Cronbach's alpha. RESULTS: Reliability of attitude scales ranged from .59 to .73. CONCLUSIONS AND IMPLICATIONS: Attitude scales were created for the purpose of assessing youth thoughts and beliefs about the local or regional food system. Such assessments can serve as a starting point for food system educators interested in developing effective curricula on food system topics.

Adolescent↗

Label-free detection of proteins in crude cell lysate with antibody arrays by a surface plasmon resonance imaging technique.

We established a label-free method of measuring proteins in crude cell lysate using antibody arrays and surface plasmon resonance (SPR) imaging. The refractivity of the running buffer was adjusted with that of the lysate to overcome the bulk effect. The chemistries of the fabricated arrays were investigated to reduce nonspecific adsorption on the array surface. We found that the hydrophilicity of the poly(ethylene glycol) moiety and lower electrostatic charge on the surface provided a specific measurement of antigen-antibody interaction. We validated the system by measuring the expression of eight proteins in the mouse brain and comparing the results to those by conventional Western blotting. The detection limit of the antibody array was approximately 30 ng/mL in crude cell lysate, on the same order as that of previous SPR research. The system enabled quick, label-free, and high-throughput analysis of abundant proteins with minimal sample volume ( approximately 200 muL). It is expected that our SPR antibody array will be applicable for direct protein expression profiling of cell lysate, as well as for cell phenotyping, food analysis, discovery of new biomarkers, and immunological disease diagnostics.

Animals↗

[Determination of feeding value based on simple food characteristics. 1. Determination of the digestibility of green foods and their preservation products].

Regression techniques may be used to establish close relations between the crude fibre content and digestibility data of organic materials (total digestibility). It was shown from regression equations calculated for a number of feedingstuffs that the total digestibility of a feedstuff may be calculated, with a sufficient degree of accuracy, from the crude fibre content of the material. The range of variations from most of the digestibility data obtained for various feedstuffs, was clearly below the limit given (2.5%). When regression equations are used the variation limits should always be indicated for which they had been established. Differences within the different groups of feedstuffs are so small that in many instances, the same equation may generally be used for feedstuffs of the same group. Similarly, equations for calculating digestibility values may be derived on the basis of the lignin content of a material, although the range of variations will, in this case, be wider when compared with the crude fibre content. The reason for this must be seen in the greater uncertainty of the lignin method. Analogous regression equations may be found for the different conserving products of green feeds, in which certain alterations have to be made corresponding to the different methods of conserving used. On the one hand, these are accounted for by changes in the crude fibre content brought about by biochemical reactions during the conserving process on the other hand a special influence of conserving form can be observed. The content of digestible crude protein in green feeds can be established, with a high degree of accuracy, from the crude protein content of the material. The content of digestible organic matter can be calculated from the digestibility of organic matter and from the ash content of the material. The content of digestible organic matter is, in turn, closely related to the content of energetic feed units in a particular material.

Animal Feed↗

Evaluation of selective and non-selective enrichment PCR procedures for Salmonella detection.

AIMS: To compare PCR combined with enrichment media with the standard microbiological techniques (SMT) and to determine the most sensitive method for the detection of Salmonella and the identification of Salm. typhimurium (ST), Salm. enteritidis (SE), Salm. gallinarum (SG) and Salm. pullorum (SP). METHODS AND RESULTS: We analysed 87 samples from poultry using PCR and SMT, PCR being performed from non-selective (NS) and Rappaport-Vassiliadis (RV) media. PCR-NS was less sensitive than PCR-RV and SMT for the detection and identification of Salmonella. PCR-RV detected more positive samples of Salmonella sp. than SMT but both these methods showed similar sensitivity regarding the identification of Salmonella serovars. CONCLUSIONS: PCR-RV was more sensitive and decreased the time necessary to detect and identify Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: PCR-RV is a powerful tool for the rapid and accurate detection and identification of Salmonella and can be implemented in diagnostic and food analysis laboratories.

Animals↗

Simultaneous detection of enteric viruses by multiplex real-time RT-PCR.

A multiplex real-time RT-PCR protocol for the simultaneous detection of noroviruses ("Norwalk-like viruses") of genogroups I and II, human astroviruses and enteroviruses is described. The protocol was developed and evaluated using the LightCycler and corresponding SYBR Green reagents. New primers were designed within conserved genome regions to optimize the detection range of virus subtypes of each genus. To enable the development of a multiplex PCR assay within one tube (capillary), similar mastermix- and cycling-conditions were respected for each individual primer system. Subsequent melting curve analysis allowed the determination of possible dual-contaminations of entero- and noro- or astroviruses by the formation of dual peaks. Special care was taken to minimize the loss of sensitivity, since the detection of small viral contaminations is a crucial parameter especially for food analysis. The multiplex assay was compared successfully to the single SYBR Green assay, and revealed to be at least 10 times more sensitive than the one obtained with an endpoint PCR thermocycler protocol published previously.

Benzothiazoles↗

Determination of polycyclic aromatic hydrocarbons in smoked meat products and smoke flavouring food additives.

Formation, factors affecting concentrations, legal limits and occurrence of polycyclic aromatic hydrocarbons in smoked meat products and smoke flavour additives are briefly reviewed. The most used techniques such as thin-layer chromatography (TLC), gas chromatography (GC) and high-performance liquid chromatography (HPLC) are evaluated. Also, sample preparation, pre-separation procedures, separation and detection systems being used for determination are discussed with emphasis to latest development in applied food analysis and the chosen data regarding the concentration of polycyclic aromatic hydrocarbons in smoked meat products and smoke flavour additives are summarised.

Chromatography, Gas↗